Fibrinogen is not synthesized by human megakaryocytes

Abstract

Abstract The origin of platelet fibrinogen is controversial. It may arise from two sources: (a) exogenously by endocytosis of plasma fibrinogen, or (b) endogenously by synthesis. We explored the second possibility because we previously demonstrated that the first mechanism does occur. Fibrinogen synthesis by human megakaryocytes (MK) was investigated by in situ hybridization and the polymerase chain reaction (PCR) applied to mRNA. MK differentiating from marrow CFU-MK were cultured in suspension. In situ hybridization using the 35S alpha and beta fibrinogen chain anti-sense riboprobes was totally negative in MK in comparison with negative controls (lambda phage and alpha and beta fibrinogen chain sense riboprobes). In contrast, synthesis of fibrinogen was detected by this technique in a hepatoma cell line (HepG 2). Furthermore, mRNA for alpha and beta chains of fibrinogen was not detected by the PCR performed on mRNA from cultured MK enriched to 90% purity, by the immunomagnetic bead technique, even after Southern blotting of the amplified products. In addition, fibrinogen mRNA was undetected in marrow MK and in platelets by the same technique, whereas a specific megakaryocyte gene transcript (GPIb alpha) was easily detected. These observations demonstrate that the only mechanism responsible for the presence of fibrinogen in platelets is endocytosis of fibrinogen from plasma.